Those plasmids are very useful vectors to amplify promoters and to measure their forces using Standard Promoter Units.
The principle is easy to understand.
Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. The standard GFP tripart (E0240) is between the biobrick site, it is just between two PstI restriction sites.
This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.
| Ligation name
| Nb of colonies
| Description
| Volume gel extraction
|
| L100
| 0
| rbs-TetR <-> ECFP (RBS+LVA+Term)
| 3+6
|
| L101
| (1)
| rbs-TetR <-> GFP tripart+strong rbs
| 3+6
|
| L102
| +/- 100
| strong RBS (0,6)<-> YFP
| 5+10
|
| L103
| a lot ++
| strong RBS (0,6)<-> GFP
| 5+5
|
| L104
| a lot ++
| strong RBS (0,6)<-> LasR activator (+LVA)
| 5+4
|
| L105
| a lot ++
| strong constitutive promoter <-> ECFP
| 5+8
|
| L106
| a lot ++
| strong constitutive promoter <-> GFP tripart
| 5+8
|
| L107
| a lot ++
| strongest constitutive promoter <-> ECFP
| 5+8
|
| L108
| +/- 100 (2 dishes!?)
| strongest constitutive promoter <-> GFP tripart
| 5+8
|
| L109
| +/- 30 or a lot ++ (2 dishes !?)
| medium constitutive promoter <-> ECFP
| 4+7
|
| L110
| a lot ++
| medium constitutive promoter <-> gfp tripart
| 4+7
|
| L111
| a lot ++
| weak constitutive promoter <-> ECFP
| 3+5
|
| L112
| a lot ++
| weak constitutive promoter <-> gfp tripart
| 3+5
|
| L113
| 1
| Arac pBAD <-> ECFP
| 5+2.5
|
| L114
| 2
| Arac pBAD <-> gfp tripart
| 5+2.5
|
| L115
| a lot ++
| pLas <-> ECFP
| 7+11
|
| L116
| a lot ++
| pLas <-> gfp tripart
| 7+11
|
| L117
| +/- 30
| Double terminator <-> YFP
| 4+3
|
| L118
| +/- 30
| Double terminator <-> mRFP
| 4+2
|
| L119
| +/- 30
| Double terminator <-> LasR activator (+LVA)
| 4+2
|
| L120
| a lot ++
| tetR repressible promoter <-> ECFP
| 6+7.5
|
| L121
| a lot ++
| tetR repressible promoter <-> gfp tripart
| 6+7.5
|
| L122
| 0
| B0034rbs-LasI <-> ECFP
| 7+3.5
|
| L123
| 1
| B0034rbs-LasI <-> gfp tripart
| 7+3.5
|
| L124
| a lot ++
| strongest RBS (1) <-> GFP
| 6+5
|
| L125
| a lot ++
| strongest RBS (1) <-> YFP
| 6+10
|
| L126
| no dish found!!!
| strongest RBS (1) <-> activator (+LVA)
| 6+4
|
| L127
| no dish found!!!
| Double terminator <-> GFP
| 6+3
|
| Controls
|
|
|
|
| C1
| 0
| D110
| 3
|
| C2
| 5
| D129
| 5
|
| C3
| +/- 100
| D123
| 5
|
| C4
| +/- 20
| D103
| 5
|
| C5
| +/- 100
| Double terminator <-> GFP
| 4
|
| C6
| +/- 100
| Double terminator <-> GFP
| 3
|
| C7
| 0
| Double terminator <-> GFP
| 5
|
| C8
| +/- 20
| Double terminator <-> GFP
| 7
|
| C9
| 0
| Double terminator <-> GFP
| 4
|
| C10
| 0
| Double terminator <-> GFP
| 6
|
| C11
| 0
| Double terminator <-> GFP
| 7
|
| C12
| +/- 10
| Double terminator <-> GFP
| 6
|
| Positive control
| 155 (transformation efficiency:1.5*10^7/ug)
| ???
|
|
"
